线粒体DNA单倍型类群与HBV感染和临床结局相关性研究

发布时间：2018-07-08 10:42

本文选题：慢性乙型肝炎病毒 + 感染　；参考：《昆明医科大学》2017年硕士论文

【摘要】：[目的]以慢性HBV感染人群、H]BV自限性感染人群和健康人群为研究对象,获取其线粒体DNA控制区的遗传变异和单倍型类群信息;明确研究组间线粒体DNA变异位点和单倍型类群的分布情况,分析不同人群变异位点和单倍型类群的差异。结合临床数据,探讨候选线粒体DNA突变和单倍型类群与HBV易感性和感染结局的相关性。[方法]我们首先测定了来自云南省第二人民医院肝病内科和体检中心的234例HBV自限性感染人群的外周血的线粒体DNA控制区序列,整合本课题组前期获得的272例慢性HBV感染人群和312例健康人群线粒体DNA控制区序列进行分析。将所获得的数据和修订后的剑桥参考序列(revised Cambridge Refrence Sequence;rCRs)进行比对,我们记录了所有个体线粒体DNA控制区的种系突变信息。根据突变信息,我们参考已发表的世界人群线粒体DNA系统关系发育树,将研究对象的线粒体DNA数据划分到特定的单倍型类群中。通过X2检验,我们对慢性HBV感染人群、HBV自限性感染人群和健康人群线粒体DNA控制区突变和单倍型类群的分布频率进行统计分析。结合临床相关信息,对线粒体DNA单倍型类群的分布频率与临床特征的相关性进行了分析。[结果]1线粒体DNA控制区研究结果:通过对来自云南省第二人民医院肝病内科和体检中心的272例慢性HBV感染者、234例HBV自限性感染者和312例健康对照者外周血线粒体DNA测序,获得818份线粒体DNA控制区全长1124bp测序序列,慢性HBV感染组共计280个多态性位点,HBV自限性感染组共计272个多态性位点,健康对照组共计297个多态性位点;其中慢性HBV感染组与健康对照组重叠多态性位点共计194个,慢性HBV感染组特异位点85个;慢性HBV感染组与HBV自限性感染组重叠多态性位点共计230个,慢性HBV感染组特异性位点50个;HBV自限性感染组和健康对照组重叠多态性位点共计168个,HBV自限性感染组特异性位点104个。通过对818条序列进行了单倍型类群的划分,其中700条序列(慢性HBV感染人群231例,HBV自限性感染人群207例,健康人群262例)划分入14种单倍型类群,分别为M*、M7、M8、C、D、D4、D5、A、R9、F、F1、B、B4、B5。未归入以上14种单倍型类群者共118例(慢性HBV感染人群41例;HBV自限性感染人群27例;健康人群50例)。结合前人的数据进行主成分分析,结果显示我们研究的慢性HBV感染人群、HBV自限性感染人群和健康人群来源一致。2对mtDNA控制区突变位点进行X2检验,慢性HBV感染人群和健康人群突变位点分布的分析显示慢性HBV感染人群携带m.151CT的突变频率(2.12%)显著低于健康人群(7.37%);慢性HBV感染人群和HBV自限性感染人群的突变位点分布分析显示慢性HBV感染人群携带m.146TC、m.310CT、m.310+C、m.523-524AC/del 的突变频率(15.81%,20.22%,70.22%,41.18%)显著高于HBV自限性感染人群(8.54%,11.54%,53.85%,25.64%),慢性HBV感染人群携带m.315+C、m.16274CT突变频率(5.88%,3.30%)显著低于HBV自限性感染人群(26.92%,7.26%);对HBeAg阳性和HBeAg阴性慢性HBV感染人群的突变位点进行分析显示HBeAg阳性人群携带m.16266CT的突变频率(9.68%)明显高于HBeAg阴性人群(2.38%)。3 mtDNA单倍群类型分布频率分析,结果显示慢性HBV感染人群携带单倍群类型D5的分布频率(8.09%)明显高于健康人群(3.20%);慢性HBV感染人群携带单倍群类型D5的分布频率(8.09%)明显高于HBV自限性感染人群(3.84%);健康人与HBV自限性感染者、HBeAg阳性与HBeAg阴性慢性HBV感染者中单倍型类群分布无差异。4对携带mtDNA单倍型类群D5和非携带单倍型类群D5的慢性HBV感染人群、HBV自限性感染人群、健康人群、HBeAg阳性和HBeAg阴性慢性HBV感染人群进行一般临床特征分析,结果显示三组研究对象性别、年龄和肝功能指标无差异;5从性别层面上对男女分别进行单倍型类群分布频率的比对,发现各组间单倍型类群分布无差异。[结论]1线粒体DNA多态性位点m.151CT在健康人群中的频率显著高于慢性HBV感染者,提示该位点变异可能是慢性HBV感染的潜在保护因素,即未携带该突变的健康人群可能对HBV易感;2 多态性位点 m.146TC、m.310CT、m.310+C、m.523-524AC/del 在慢性 HBV感染人群中的频率明显高于HBV自限性感染人群,提示该变异位点可能是HBV持续感染的潜在危险因素;慢性HBV感染者携带m.315+C、m.16247CT突变频率显著低于HBV自限性感染者,提示该变异位点可能与HBV清除相关;3多态性位点m.16266CT在HBeAg阳性人群的突变频率明显高于HBeAg阴性人群,提示m.16266CT变异可能与HBV慢性感染结局不同相关。4 mtDNA单倍型类群D5可能是HBV持续感染的危险因素,且与性别、年龄肝功能无关。
[Abstract]:[Objective] to study the genetic variation and the haplotype group information of the mitochondrial DNA control area with chronic HBV infected people and H]BV self limiting infection population and healthy population, and to clarify the distribution of the mitochondrial DNA mutation and haplotype groups in the study group, and analyze the differences in the variation and haplotype groups of different populations. The correlation of the candidate mitochondrial DNA mutation and the haplotype group to the susceptibility to HBV and the infection outcome. [Methods] we first measured the mitochondrial DNA control region of the peripheral blood from 234 cases of HBV self limited infection from the second people's Hospital of Yunnan province. The sequence of the mitochondrial DNA control region of 272 chronic HBV infected people and 312 healthy people was analyzed. The data obtained were compared with the revised Cambridge reference sequence (revised Cambridge Refrence Sequence; rCRs). We recorded the mutation information of the mitochondrial DNA control area of all individuals. The mitochondrial DNA system of the world population was published, and the mitochondrial DNA data of the subjects were divided into a specific haplotype group. Through X2 test, we analyzed the distribution frequency of the mitochondrial DNA control region and the haplotype groups in the chronic HBV infected population, the HBV self limiting infection population and the healthy population. Combined with clinical information, the correlation between the distribution frequency of mitochondrial DNA haplotype groups and clinical characteristics was analyzed. [results]1 mitochondrial DNA control area study results: 272 cases of chronic HBV infection, 234 cases of HBV self limiting infection and 312 healthy pairs of HBV from the liver disease internal medicine and physical examination center of the second people's Hospital of Yunnan province. In the peripheral blood mitochondrial DNA sequencing, 818 mitochondrial DNA control region full length 1124bp sequencing sequences, 280 polymorphic loci in the chronic HBV infection group, 272 polymorphic loci in the HBV self limiting infection group, and 297 polymorphic loci in the healthy control group, of which the chronic HBV infection group and the healthy control group were polymorphic loci in total. 194, 85 special sites in chronic HBV infection group, 230 overlapped polymorphic loci in chronic HBV infection group and HBV self limiting infection group, 50 specific loci in chronic HBV infection group, 168 polymorphic loci in HBV self limiting infection group and healthy control group, 104 specific loci in the self limited infection group. 818 sequence entered into 818 sequences. 700 sequences (231 cases of chronic HBV infection, 207 cases of HBV self limiting infection, 262 healthy people) were divided into 14 haplotypes, including M*, M7, M8, C, D, D4, D5, A, R9, 118 cases (41 cases of chronic infection). 27 people and 50 healthy people. Combined with previous data, principal component analysis showed that we studied chronic HBV infected people, HBV self limiting infection population and healthy population origin consistent.2 to X2 test of mutation site in mtDNA control area. Analysis of chronic HBV infection and distribution of mutation sites in healthy population showed chronic HBV The mutation frequency (2.12%) of the infected people (2.12%) was significantly lower than that of the healthy population (7.37%), and the distribution analysis of the mutation sites in the chronic HBV infected people and the HBV self limited infection population showed that the frequency of mutation frequency of m.146TC, m.310CT, m.310+C, m.523-524AC/del (15.81%, 20.22%, 70.22%, 41.18%) of chronic HBV infected people (15.81%, 20.22%, 70.22%, 41.18%) was significantly higher than that of the HBV self limiting sex Infected people (8.54%, 11.54%, 53.85%, 25.64%), chronic HBV infected people carrying m.315+C, m.16274CT mutation frequency (5.88%, 3.30%) significantly lower than the HBV self limited infection population (26.92%, 7.26%); HBeAg positive and HBeAg negative chronic HBV infected population of the mutation site analysis showed that HBeAg positive people carry the mutation frequency of m.16266CT (9.68%) Ming (9.68%) The frequency analysis of the single group type distribution of.3 mtDNA was significantly higher than that of HBeAg negative group (2.38%). The results showed that the frequency of the distribution of unhaploid group D5 in chronic HBV infected people (8.09%) was significantly higher than that of the healthy population (3.20%), and the frequency of the distribution of unhaploid group D5 in chronic HBV infected people (8.09%) was significantly higher than that of the HBV self restricted population (3.84%). There was no difference in the distribution of haplotype groups in patients with HBV self limiting infection, HBeAg positive and HBeAg negative chronic HBV infection,.4 for chronic HBV infected people carrying mtDNA haplotype group D5 and non haplotype group D5, HBV self limiting infection population, healthy population, HBeAg positive and HBeAg negative chronic infection population. The results showed that there was no difference in sex, age and liver function between the three groups. 5 the distribution frequencies of haplotype groups were compared between men and women at the gender level, and there was no difference in the distribution of haplotype groups in each group. [conclusion]1 mitochondrial DNA polymorphism site m.151CT was significantly higher in healthy people than in chronic HBV sense. The mutant may be a potential protective factor for chronic HBV infection, that is, healthy people who do not carry the mutation may be susceptible to HBV, and 2 polymorphic loci, m.146TC, m.310CT, m.310+C, and m.523-524AC/del are significantly higher in chronic HBV infected people than in HBV self limiting infections, suggesting that the heterotopic point may be a HBV holding. The potential risk factors for continued infection; chronic HBV infected people carrying m.315+C, the mutation frequency of m.16247CT was significantly lower than that of HBV self limiting infection, suggesting that the heterotopic point may be associated with HBV clearance; 3 the mutation frequency of m.16266CT in HBeAg positive population is significantly higher than that of HBeAg negative population, suggesting that m.16266CT variation may be associated with HBV chronic infection. .4 mtDNA haplotype D5 may be a risk factor for persistent HBV infection and is not associated with gender, age and liver function.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.62

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